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从果蝇头部制备染色质和进行染色质免疫沉淀。

Chromatin Preparation and Chromatin Immunoprecipitation from Drosophila Heads.

机构信息

CNR Institute of Molecular Genetics "Luigi-Luca Cavalli-Sforza", Unit of Bologna, Bologna, Italy.

IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy.

出版信息

Methods Mol Biol. 2023;2655:19-30. doi: 10.1007/978-1-0716-3143-0_2.

Abstract

Chromatin immunoprecipitation (ChIP) is a widely used method to map protein-DNA interactions in vivo. Formaldehyde cross-linked chromatin is fragmented, and the protein of interest is immunoprecipitated using a specific antibody. The co-immunoprecipitated DNA is then purified and analyzed by quantitative PCR (ChIP-qPCR) or next-generation sequencing (ChIP-seq). Therefore, from the amount of DNA recovered, it can be inferred the localization and abundance of the target protein at specific loci or throughout the entire genome. This protocol describes how to perform ChIP from Drosophila adult fly heads.

摘要

染色质免疫沉淀(ChIP)是一种广泛用于在体内绘制蛋白质-DNA 相互作用的方法。甲醛交联的染色质被片段化,然后使用特定的抗体免疫沉淀感兴趣的蛋白质。随后,通过定量 PCR(ChIP-qPCR)或下一代测序(ChIP-seq)对共免疫沉淀的 DNA 进行纯化和分析。因此,从回收的 DNA 量可以推断出目标蛋白在特定基因座或整个基因组中的定位和丰度。本协议描述了如何从果蝇成虫头部进行 ChIP。

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